Table 2. Tumorigenicity testing by subcutaneous transplantation of hiPSC-derived RPE into NOG mice.

hiPSC cell line	cell form	min.dose fortumor formation	weeks to observeTumor (first to last)	numberof mice	Log10TPD50
201B7	Cell suspension in Matrigel	1×101 cells	5–40	30	2.12
RPE cell line	cell form	number of cells transplanted	monitor period	number of mice	tumor formation
59-G3(1)	RPE cell suspension in Matrigel	1×106 cells	26–84 weeks	9	none
K21-G18	RPE cell suspension in Matrigel	1×106 cells	26–74 weeks	8	none
101-G25	RPE cell suspension in Matrigel	1×106 cells	23–70 weeks	10	none
59-G3(1)	RPE cell sheet in Matrigel	1×106 cells	28–85 weeks	5	none
K21-G18	RPE cell sheet in Matrigel	1×106 cells	13–79 weeks	5	none
101-G25	RPE cell sheet in Matrigel	1×106 cells	23–79 weeks	5	none
primary RPE	Cell suspension in Matrigel	1×106 cells	52 weeks	3	none
primary RPE	Cell suspension w/o Matrigel	1×106 cells	52 weeks	2	none
59-G3(2)	RPE cell sheet in Matrigel	1×106 cells	26–50 weeks	3	none
RNT10	RPE cell sheet in Matrigel	1×106 cells	26–46 weeks	3	none
RNT9	RPE cell sheet in Matrigel	1×106 cells	26–38 weeks	3	none
101-EV3	RPE cell suspension in Matrigel	1×106 cells	39 weeks	5	none
K11-EV9	RPE cell suspension in Matrigel	1×106 cells	39 weeks	3	none
K21-EV15	RPE cell suspension w/o Matrigel	1×106 cells	39 weeks	4	none
K11-EV9	RPE cell suspension w/o Matrigel	1×106 cells	39 weeks	2	none

Log₁₀TPD₅₀ value for hiPSC 201B7 determined by subcutaneously transplanting cells in Matrigel into NOG was calculated by the Trimmed Spearman-Karber method (upper panel). Tumor formation from 1×10⁶ hiPSC-derived RPE cells prior to making RPE sheets (cell suspension) or after making RPE sheets (cell sheet) transplanted subcutaneously in various conditions into NOG mice. Animals were monitored for 13–85 weeks (lower panel).